Transient Expression of Tomato miR482a Targets an NBS-LRR mRNA in N. benthamiana Resulting in RDR6-Dependant Phased siRNA Biogenesis.
(A) Targeting of Nicotiana EU713768.1 NBS-LRR by miR482a. The alignment shows the target site of miR482a in a tobacco NBS-LRR mRNA, and the predicted sequences of phased secondary siRNAs are shown at an adjacent site D1. Δ indicates modified sequences underlined in the miR482aΔ construct.
(B) RNA gel blot analysis of miR482a and D1 siRNA in Agrobacterium tumefaciens infiltrated zones of N. benthamiana wild-type (WT) and RDR6i lines. Analyses of miR168 and U6 RNAs are included as controls. M, decade marker indicating a 20-nucleotide RNA.
(C) U713768.1 mRNA accumulation 3 d after inoculation by quantitative PCR. See Methods for construct design. Error bars indicate sd (n = 3), and asterisks indicate a significant difference from corresponding control samples (t test, P value < 0.05).